紫外诱变选育巴弗洛霉素A1高产菌株及其培养基优化  

The High-yield Bafilomycin A1 Strain Obtained from UV Mutagenesis and Its Medium Optimization

作　　者：陈海敏 孙菲[2] 袁源 吴佳雯 江红[2] 周剑 CHEN Hai-min;SUN Fei;YUAN Yuan;WU Jia-wen;JIANG Hong;ZHOU Jian(School of Pharmacy,Fujian Medical University,Fuzhou 350122;Fujian Key Laboratory of Screening for Novel Microbial Products,Fujian Institute of Microbiology,Fuzhou 350007)

机构地区：[1]福建医科大学药学院,福州350122 [2]福建省微生物研究所、福建省新药(微生物)筛选重点实验室,福州350007

出　　处：《生物技术通报》2025年第3期51-61,共11页Biotechnology Bulletin

基　　金：福建省属公益类科研院所基本科研专项(2024R1005007,2023R1005004)。

摘　　要：【目的】巴弗洛霉素是由灰褐色链霉菌(Streptomyces griseobrunneus)发酵产生的一类大环内酯化合物,对多种植物真菌具有拮抗作用,其代表性化合物为巴弗洛霉素A1(Baf A1)。为了进一步提升链霉菌(Streptomyces sp.)FIM-B0711合成Baf A1的发酵产量,降低生产成本,以其为出发菌株选育Baf A1高产菌株,并对其发酵工艺进行优化。【方法】采用紫外诱变选育高产突变株,单因素试验、最陡爬坡试验和响应面法进行摇瓶发酵工艺优化。【结果】紫外诱变菌株UV-21的Baf A1产量达到397.99 mg/L,较原始菌株提高了23.60%。单因素试验表明,该菌株发酵的最佳碳源、氮源和无机盐分别为麦芽糊精、大豆蛋白胨和碳酸钙,最佳培养条件为pH 7.0、接种量3%(体积分数)、装液量30 mL/250 mL、发酵培养时间96 h。最陡爬坡试验表明,在麦芽糊精45 g/L、大豆蛋白胨15 g/L、碳酸钙0.5 g/L时相对效价达到最高,为147.18%。响应面优化试验表明,该菌株的最佳发酵工艺:麦芽糊精45.98 g/L、大豆蛋白胨14.96 g/L、碳酸钙0.48 g/L、缬氨酸3 g/L,初始pH 7.0、接种量3%(体积分数)、装液量30 mL/250 mL、发酵培养时间96 h。优化后高产菌株UV-21摇瓶发酵产量为627.58 mg/L,较初始工艺提高了57.69%。【结论】基于紫外诱变及培养基的响应面优化试验获得1株Baf A1产量为627.58 mg/L的高产菌株,较原始菌株FIM-B0711产量提高了57.69%。【Objective】Bafilomycins are a class of macrolide compounds produced by the fermentation of Streptomyces griseobrunneus,which have antagonistic effect on a variety of plant fungi,and its representative compound is bafilomycin A1(Baf A1).In order to further enhance the yield of Baf A1 in Streptomyces sp.FIM-B0711 and reduce the production cost,the high-yielding strain of Baf A1 was selected and its fermentation process was optimized.【Method】High-yield mutant strains were generated through ultraviolet mutagenesis,followed by optimization of the fermentation process using single factor experiment,the steepest ascent experiment,and response surface methodology.【Result】The Baf A1 yield of the UV-21 strain reached 397.99 mg/L,representing a 23.60%increase compared to the original strain.Single factor experiment identified that maltodextrin,soybean peptone,and calcium carbonate were as the optimal carbon source,nitrogen source,and inorganic salt for strain UV-21,respectively.The optimal culture conditions were determined to be pH 7.0,an inoculum concentration of 3%(volume fraction),a liquid volume of 30 mL/250 mL,and a fermentation time of 96 h.The steepest ascent experiment revealed that the highest relative titer of 147.18%was achieved with a carbon source concentration of 45 g/L,nitrogen source concentration of 15 g/L,and inorganic salt concentration of 0.5 g/L.The response surface optimization demonstrated that the optimal fermentation process for the strain involved maltodextrin at 45.98 g/L,soybean peptone at 14.96 g/L,calcium carbonate at 0.48 g/L,valine at 3 g/L,an initial pH of 7.0,an inoculum concentration of 3%(volume fraction),a liquid volume of 30 mL/250 mL,and a fermentation time of 96 h.Under these optimized conditions,the fermentation titer reached 627.58 mg/L,a 57.69%increase compared to the initial process.【Conclusion】A high-yield Baf A1 strain with 627.58 mg/L is obtained based on ultraviolet mutagenesis and medium response surface optimization experiment,which increases the yield by 57

关 键 词：巴弗洛霉素 链霉菌 响应面试验 发酵优化 紫外诱变 培养条件 工艺参数 

分 类 号：TS2[轻工技术与工程—食品科学与工程]